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Targeted viral vector transduction of relaxin-3 neurons in the rat nucleus incertus using a novel cell-type specific promoter
انتقال بردار ویروسی هدفمند از سلولهای عصبی ریلسین -3 در هسته موش با استفاده از یک پروموتر خاص از نوع سلول جدید-2020 Modern neuroscience utilizes transgenic techniques extensively to study the activity and function of brain neural
networks. A key feature of this approach is its compatibility with molecular methods for selective transgene
expression in neuronal circuits of interest. Until now, such targeted transgenic approaches have not been applied
to the extensive circuitry involving the neuropeptide, relaxin-3. Pharmacological and gene knock-out studies
have revealed relaxin-3 signalling modulates interrelated behaviours and cognitive processes, including stress
and anxiety, food and alcohol consumption, and spatial and social memory, highlighting the potential of this
system as a therapeutic target. In the present study, we aimed to identify a promoter sequence capable of
regulating cell-type specific transgene expression from an adeno-associated viral (AAV) vector in relaxin-3
neurons of the rat nucleus incertus (NI). In parallel to relaxin-3 promoter sequences, we also tested an AAV vector
containing promoter elements for the tropomyosin receptor kinase A (TrkA) gene, as TrkA is co-expressed with
relaxin-3 in rat NI neurons. Stereotaxic injection of an mCherry-expressing AAV vector revealed widespread nonspecific
TrkA promoter (880 bp) activity in and adjacent to the NI at 8 weeks post-treatment. In contrast,
mCherry expression was successfully restricted to relaxin-3 NI neurons with 98% specificity using a 1736 bp
relaxin-3 promoter. In addition to detailed anatomical mapping of NI relaxin-3 networks, illustrated here in
association with GABAergic medial septum neurons, this method for targeted transgene delivery offers a versatile
tool for ongoing preclinical studies of relaxin-3 circuitry. Keywords: Adeno-associated viral (AAV) vector | Cell-type specific promoter | Medial septum | Nucleus incertus | Relaxin-3 | Tropomyosin receptor kinase A (TrkA) |
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